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Öğe Glycoside oleandrin downregulates toll-like receptor pathway genes and associated miRNAs in human melanoma cells(Elsevier, 2022) Güneş, Canan Eroğlu; Çelik, Fatma Seçer; Seçme, Mücahit; Elmas, Levent; Dodurga, Yavuz; Kurar, ErcanMelanoma accounts for the majority of skin cancer-related deaths. Nerium oleander is a plant known to be toxic and consumed due to the cardiac glycosides it contains. Oleandrin is a cardiac glycoside obtained from of N. oleander. Beside capable of inhibiting proliferation and metastasis of cancer cells, cardiac glycoside derivative compounds cause cardiovascular side effects. Because of cardiovascular toxicity of clinically used cardiac gly-cosides, it is necessary to investigate cardiac glycoside derivative compounds capable of inhibiting proliferation and metastasis of cancer cells.It is known that oleandrin has anticarcinogenic effects in other cancers. Previous studies have shown that toll -like receptors (TLRs) and their related microRNAs (miRNAs) are associated with cancer. Therefore, aim was to investigate the effect of oleandrin on genes and miRNAs associated with TLRs in A375 melanoma cells in this study. The effects of oleandrin on cell viability, cytokines, apoptosis were evaluated using XTT, ELISA and TUNEL analyses, respectively. The effect of oleandrin on expression of TLR genes and 5 associated miRNAs in A375 cells has been determined by qRT-PCR. In addition, the levels of MyD88, TLR2 and TLR4 proteins were analyzed by western blot method.ELISA indicated that oleandrin treatment (47 nM at 48 h) reduced the level of proinflammatory cytokine IFNG. TUNEL analysis showed that apoptosis rate was significantly increased in the oleandrin dose group. According to qRT-PCR results, there was a significant decrease in IRAK1, IRAK4, MyD88, TLR2-TLR7 and TRAF3 expressions in the oleandrin treated group compared to the control (untreated cell). Also, a significant decrease in TLR4 protein expression has been observed. In addition, oleandrin significantly downregulated the levels of hsa-miRNA-146a-5p and hsa-miRNA-21-5p.In conclusion, it has been observed that oleandrin has an effect on TLR pathway-related genes and miRNAs in melanoma cells. We show that TLRs pathways and hsa-miR-146a-5p and hsa-miR-21-5p can participate in the oleandrin molecular mechanism of action.Öğe Impact of cisplatin on Kasumi-1 leukemia cell line: gene expression and DNA damage(Pamukkale University, 2025) Dodurga, Yavuz; Seçme, Mücahit; Elmas, Levent; Demirkıran, Nazlı; Sağ, Sevda; Pala, Ulviye; Akdağ, ZelihaPurpose: Leukemia is a type of cancer caused by the uncontrolled proliferation of blood cells. The purpose of this study was to investigate the effects of cisplatin (CIS), a chemotherapeutic agent used in the treatment of leukemia, on the Kasumi-1 leukemia cell line. Materials and methods: The study measured the effect of CIS on Kasumi-1 cells by calculating IC50 values for cell viability. The mRNA expression levels of apoptosis and cell cycle-related genes were then assessed using Real-Time PCR. In addition, the effects of CIS on DNA damage were investigated using the comet assay. Results: Significant changes in apoptosis and cell cycle-related genes were observed in CIS-treated groups. These included alterations in the mRNA levels of p53, BCL-2, CHECK 1, CDC25C, CDK 6, URG4/URGCP, GADD45A, CCND1, GADD45G, and ATM genes. Comet analysis confirmed CIS's effects on DNA damage. Conclusion: This study aimed to better understand how CIS affects genetic mechanisms in leukemia cells and provide new insights into leukemia treatment. The findings will help us better understand the role of CIS in leukemia treatment and will serve as a valuable reference for future research. © 2025, Pamukkale University. All rights reserved.Öğe Investigation of the apoptotic and cell cycle effects of sorafenib and doxorubicin on URG4/URGCP in leukemia cells(Pamukkale University, 2024) Dodurga, Yavuz; Elmas, Levent; Seçme, Mücahit; Demirkıran, Nazlı; Avcı, Çığır Biray; Bağcı, Gülseren; Sağ, SevdaPurpose: The aim of this study is to investigate the effects of anticancer drugs such as Sorafenib (SOR) and Doxorubicin (DOX) on URG4/URGCP mRNA levels in K562 and HL-60 leukemia cells, elucidating their effects on apoptosis and cell cycle. The effects of these drugs on apoptosis and the cell cycle in leukemia cells have been explored. This research aims to understand the cellular effects of drugs used in leukemia treatment and contribute valuable insights to the drug development processes in leukemia therapy. Materials and methods: DOX and SOR were evaluated for their IC50 values in K562 and HL-60 cell lines using the CellTiter-Glo assay (Promega, USA), based on ATP measurement. Total RNA isolation was performed using Trizol reagent in both control and dose groups of each treated cell line. Following RNA isolation, cDNAs were synthesized using the "Transcriptor High Fidelity cDNA Synthesis Kit". Subsequently, changes in mRNA expression levels were examined using specific primers for URG4/URGCP, Casp-3, Casp-8, Casp-9, FADD, DR4, TRADD, CCDN1, CDK4, CDK6, PTEN, P53, and Rel-A genes. Results: In the groups treated with Sorafenib, the IC50 dose for HL-60 cell line was calculated as 40 µM at the 24th hour, and for K562 cell line, it was calculated as 40 µM at the 48th hour. In the groups treated with Doxorubicin, the IC50 doses were calculated as 50 µM at the 48th hour for HL-60 cell line, and as 50 µM at the 72nd hour for K562 cell line. Significant increases were observed in the mRNA expression levels of Casp-8, Casp-9, TRADD, DR4, Rel A, and FADD genes in the groups treated with SOR, while a decrease was observed in the mRNA expression levels of URG4/URGCP, CCDN1, CDK4, and CDK6 genes. In the groups treated with DOX, significant increases were observed in the fold changes of Casp-3, Casp-8, P53, and PTEN genes. However, a significant decrease in mRNA expression levels was observed in URG4/URGCP, CCDN1, and CDK4 genes. Conclusion: As a result, it has been demonstrated that both SOR and DOX may play a role in regulating the mRNA expressions of URG4/URGCP, Casp-3, Casp-8, Casp-9, CDK6, CDK4, CCND1, P53, PTEN, TRADD, DR4, Rel A, and FADD genes in HL-60 and K562 cells. © 2024, Pamukkale University. All rights reserved.Öğe Potential effects of parietin on apoptosis and cell cycle related genes in SH-SY5Y neuroblastoma cells(Pamukkale University, 2024) Dodurga, Yavuz; Seçme, Mücahit; Elmas, Levent; Gündoğdu, Gülşah; Çekin, Ayşe; Günel, Nur SelviPurpose: Ingredients obtained from natural products have been used in cancer treatments for years. High diversity and non-toxicity compared to chemotherapeutic agents are the main reasons for their preference. Lichens having potential for treatment of cancer consist of fungus and 1-2 species of algae. Under the name of lichen substances, many of them have also been synthesized as specific substances. The secondary metabolites in lichens are generally insoluble in water, and have many biological activities such as antiviral, antitumor, antibacterial, and antioxidant; they store in the fungal cell or on the surface of the hyphae and can only be extracted with organic solvents. Parietin extracted from lichen species such as xanthoria parietina is an anthraquinone pigment and a secondary metabolite. In our study, the effects of parietin on cytotoxicity, gene expression, migration, invasion, and colony formation in neuroblastoma cells treated with parietin were investigated. SH-SY5Y cell line without parietin was used as the control group. Materials and methods: The IC50 value of the parietin was determined using XTT assay. The total RNA extractions were performed from the cells using the Tri-Reagent kit. The expressions of BAX, CASPASE3, CASPASE8, CASPASE9, P53, PUMA, NOXA, TIMP1, TIMP2, BCL2, BCL-XL, CASPASE10, BID, CYCLIND1, CDK6, P21, MMP2, MMP9, TRADD and FADD genes were investigated by Lightcycler 480 (Roche) using SYBR Green dye. Migration analysis of the control and the dose group cells were performed in accordance with the Wound-healing assay protocol. Invasion activities were determined using the “Invasion Chamber” (BD Biosciences) protocol. Colonies were treated with crystal violet and observed under the light microscope. Results: The IC50 value of the parietin used for 48-hour treatment on the cells was determined as 35 µM. It was found that the expression levels of BCL-XL, BCL-2, MMP2, MMP9, P21, and CYCLIN D1 mRNA were downregulated, and it was also shown to be upregulated the expression levels of CASPASE3, CASPASE9, BAX, P53, PUMA, and NOXA to be upregulated. It was determined that parietin suppressed both cell invasion and migration, and colony formation in the neuroblastoma cells. Conclusions: Thus, it can be possible parietin to be used as an alternative, complementary, and supportive agent together with the other drugs in the treatment of neuroblastoma. However, more comprehensive studies supporting these significant effects of parietin will increase its potential in the application. © 2024, Pamukkale University. All rights reserved.Öğe The role of ERK-1 and ERK-2 gene polymorphisms in PCOS pathogenesis(Bmc, 2022) Guney, Gurhan; Taskin, Mine Islimye; Sener, Nazli; Tolu, Ezgi; Dodurga, Yavuz; Elmas, Levent; Cetin, OrkunBackground Ovulation is regulated by extracellular signal-regulated kinase-1 (ERK-1) and ERK-2 signaling mechanisms, and ERK-1/2 kinases modulates the function of most of the LH-regulated genes. Defective ERK kinase signaling that is secondary to a genetic problem contributes to both ovulatory dysfunction and metabolic problems in polycystic ovary syndrome (PCOS). We planned to investigate ERK-1 and ERK-2 gene polymorphisms in PCOS for the first time in the Turkish population. Methods One hundred two PCOS patients and 102 healthy controls were recruited for this patient control study. HOMA-IR, Ferriman-Gallwey score (FGS), waist-to-hip ratio (WHR), and body mass index (BMI) were assessed. Lipid profile levels, CRP, and total testosterone were determined. ERK-2 rs2276008 (G > C) and ERK-1 rs11865228 (G > A) SNPs were analyzed with a real-time PCR system. Results ERK-1 and ERK-2 genotypes were found to differ between the PCOS and control groups. In patients with PCOS, ERK-1 GA and ERK-2 GC genotypes were different in terms of BMI, FGS, HOMA-IR, CRP, total testosterone, and total cholesterol levels. Conclusions ERK-1 and ERK-2 genes are involved in PCOS pathogenesis. BMI, FGS, HOMA-IR, and CRP levels are related to the heterozygote polymorphic types of ERK-1 and ERK-2 genes.
